DNA甲基转移酶抑制剂5-杂氮-2'-脱氧胞苷对人鼻咽癌细胞株CNE2的影响

Effect of DNA Methyltransferase 5-aza-2'-deoxycytidine on Proliferation of Human Nasopharyngeal Carcinoma Cell Line

目的探讨DNA甲基转移酶抑制剂5-aza-2dC对人低分化鼻咽鳞癌细胞株CNE2的影响。方法应用10-7、10-6、10-5、10-4mol/L浓度的5-aza-2dC,处理人低分化鼻咽鳞癌细胞株CNE2 12、24、36、48、72 h后,应用MTT法测定CNE2细胞株的生存情况,应用流式细胞仪检测细胞凋亡情况。用甲基化特异性PCR检测药物处理前后死亡相关蛋白激酶(DAPK)启动子甲基化状态。结果同一作用时间不同药物浓度组间,细胞存活率差异有统计学意义(P<0.05),且作用72 h对人低分化鼻咽鳞癌细胞株CNE2生长抑制作用最强。流式细胞仪检测结果显示:10-4mol/L 5-aza-2dC诱导细胞凋亡最明显(P<0.01)。甲基化特异性PCR显示鼻咽癌细胞株中,5-aza-2dC可成功逆转DAPK基因启动子高甲基化状态。结论 5-aza-2dC对人低分化鼻咽鳞癌细胞株CNE2生长有抑制作用,且呈浓度和时间依赖性。

Objective To explore the effect of the DNA Methyltransferase 5-aza-2′-deoxycytidine（5-aza-2dC） on human nasopharyngeal carcinoma cell line（CNE2）.Methods CNE2 were treated by 5-aza-2dC with 10-7,10-6,10-5,10-4 mol/L at 12,24,36,48,72 h,respectively.The growth rate of the cells was detected by MTT assay and cell apoptosis were analyzed by flow cytometry method.Methylation specific PCR was used to detect methylation status of death-associated protein kinase（DAPK） gene promotor before and after drug treatment.Results The differentiation of the cells survival rate was significant in statistics（P〈0.05）.And inhibitory peaks were reached at 72 hours in each concentration groups.The peak apoptosis of the cells by 5-aza-2dC in 10-4mol/L was detected by flow cytometry.The results was consistent with the MTT assay.Methylation specific PCR showed that high methylation state of DAPK gene promoter in CNE2 could be inverted into its non-methylation state by 5-aza-2dC.Conclusion The proliferation of CNE2 cells was inhibited by 5-aza-2dC in a time-and-concentration-dependent manner.